LOCUS       sequence-342219-        2871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP expression under the control of E. coli clpB promoter engineered
            with riboregulators.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2871)
  AUTHORS   Saltepe B, Bozkurt EU, Haciosmanoglu N, Seker UOS
  TITLE     Genetic Circuits To Detect Nanomaterial Triggered Toxicity through 
            Engineered Heat Shock Response Mechanism.
  JOURNAL   ACS Synth Biol. 2019 Oct 18;8(10):2404-2417. doi: 
            10.1021/acssynbio.9b00291. Epub 2019 Oct 2.
  PUBMED    31536326
REFERENCE   2  (bases 1 to 2871)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.9b00291"; journalName: "ACS Synth Biol"; date: 
            "2019-10-18- 18"; volume: "8"; issue: "10"; pages: "2404-2417"
COMMENT     Sequence Label: PclpB riboswitch GFP pZa
FEATURES             Location/Qualifiers
     source          1..2871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     435..455
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      516..602
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(637..654)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(771..1316)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(851..870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1430..1524)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1545..2204)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2119..2138
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2205..2307)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      2460..2546
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2635..2646
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2653..2871,1..495)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2683..2711)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 tacccagatc atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta
       61 caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag
      121 tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat
      181 ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt atacatcatg
      241 gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat
      301 ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
      361 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
      421 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
      481 gatgaactat acaaataaaa gcttacgcgt ggcatcaaat aaaacgaaag gctcagtcga
      541 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      601 atccgccgcc ctagacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc
      661 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
      721 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
      781 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
      841 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
      901 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
      961 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     1021 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     1081 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     1141 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     1201 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca
     1261 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     1321 atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg
     1381 tagcacctga agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa
     1441 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     1501 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
     1561 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     1621 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     1681 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     1741 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     1801 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     1861 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     1921 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     1981 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     2041 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     2101 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     2161 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     2221 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     2281 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacgtcctgc
     2341 agcttgaata attgagggat gacctcattt aatctccgtc gacacccaaa tccaggaggt
     2401 gattggtagt ggtggttaat gaaaattaac ttactactac catatatcaa gcttggcatc
     2461 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     2521 tgaacgctct cctgagtagg acaaatccgc cgccctagac tcgagcttga ataattgagg
     2581 gatgacctca tttaatctcc ggatcctacc attcacctct tggatttggg tattaaagag
     2641 gagaaaggta ccatgcgtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     2701 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     2761 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     2821 tggccaacac ttgtcactac tttcggttat ggtgttcaat gctttgcgag a
//
